The present invention relates to the detection of Lyme disease, and particularly to the clinically reliable examination and detection of Lyme disease in instances of seronegativity in mammals and particularly in humans.
Seronegativity is one of the major obstacles to diagnosis of Lyme disease as well as a common unexplained feature of the disease. Lyme disease, etiologically linked to the spirochete, Borrelia burgdorferi, may present acutely or develop insidiously. Its manifestations are protean and may specifically involve the musculoskeletal, cardiovascular, cutaneous, and nervous systems. Some cases begin with a characteristic cutaneous lesion, erythema chronicum migrans (ECM), following a deer tick bite. However, many others are unaware of a tick bite or skin lesion, making diagnosis difficult, yet develop features of Lyme disease. Diagnosis is also complicated when symptomatic individuals are negative for Borrelia burgdorferi antibody despite sensitive serological assays. Untreated patients as well as patients who have received early antibiotic therapy may remain seronegative even when symptoms of infection persist.
In general, antibody may not be detected following exposure to an infectious agent either because it is bound or sequestered, and therefore hidden from conventional probes or because it is not secreted by B cells. Dattwyler et. al. have demonstrated that in Lyme disease, T cell reactivity towards Borrelia burgdorferi occurs even in the absence of significant specific antibody as detected by a sensitive ELISA (Dattwyler et al., N. ENGL. J. MED. (1988), 319:1441-1446).
Because of the inability to offer reliable detection of antibodies to Borrelia burgdorferi, and owing to the lack of predictability in the manifestation of visible symptoms of the disease such as ECM, a large population of those afflicted with the disease do not receive timely or adequate treatment and thereby risk far more serious consequences such as the development of a chronic condition and the more serious sequelae such as meningitis, chronic radiculoneuropathies, fatigue, cognitive problems, pericarditis, or arthritis. Likewise, the recent and dramatic increase in the incidence of Lyme disease and the corresponding lack of consistent and early diagnoses have severely compromised the ability to successfully treat this disease. A need therefore exists for a reliable procedure and associated materials which will facilitate the early and consistent diagnosis of the disease and permit the aggressive therapeutic protocols presently in existence as well as those under development, to be administered in timely fashion to arrest the progress of the disease and substantially reduce, if not eliminate, its continued progress and activity.